豌豆根胞质V_1-ATPase的鉴定(英文)

Characterization of Cytosolic V_1-ATPase in Pea Root

利用免疫印迹、免疫电镜和ATP水解活性的测定对豌豆 (PisumsativumL .)根细胞胞质中V1_ATPase复合物的存在进行鉴定。用兔抗绿豆V_typeH+ _ATPase的A、B亚基的抗体进行的immuno_blotting和胶体金电镜结果都表明 ,胞质中存在有A、B亚基。活性测定结果进一步表明胞质具有ATP水解活性。这些结果说明豌豆根胞质具有有活性的V1_ATPase复合物。这是首次直接证明植物中有胞质V1_ATPase的存在。

Cytosolic V-1-ATPase complex in pea (Pistum sativum L.) root cells was characterized by immunoblotting, immuno-electron microscopy and measurement of ATP hydrolysis activity. Using antibodies against the subunits A and B of V-type H+-ATPase (V-ATPase) from mung bean as primary antibodies, both immuno-blotting and immuno-electron microscopy showed that subunit A and subunit B existed in the cytosol. Measurement of ATPase activity further showed that cytosolic proteins had NO3--sensitive ATP hydrolysis activity. All these suggested that V-1-ATPase complex occurred in the cytosol. This is the direct proof that V-1-ATPase occurs in the cytosol of the plant cells for the first time.